榄香烯联合热疗对Raji细胞增殖的影响及其机制探讨

Effects of elemene combined with hyperthermia on proliferation of Raji cells and its mechanisms

目的:观察榄香烯(elemene,ELE)联合热疗(hyperthermia,HTM)对人Burkitt's淋巴瘤Raji细胞增殖及DNA拓扑异构酶Ⅰ和Ⅱ(TOPOⅠ,TOPOⅡ)表达的影响。方法:采用噻唑蓝还原法(MTT)观察ELE联合HTM对Raji细胞增殖的影响;流式细胞术(FCM)检测细胞周期;RT-PCR法检测细胞TOPOⅠ和TOPOⅡmRNA表达;Western Blot法检测细胞TOPOⅠ和TOPOⅡ蛋白表达。结果:与单纯HTM和单纯ELE比较,ELE联合HTM对Raji细胞增殖的抑制作用增强(P<0.05),呈时间和剂量依赖性,而ELE联合HTM作用Raji细胞24h、48h和72h后,IC50值(93μg/ml、68μg/ml和41μg/ml)小于单纯ELE IC50值(116μg/ml、84μg/ml和60μg/ml);ELE联合HTM阻滞Raji细胞于S期[(35.40±1.27)%、(42.38±2.50)%VS(48.88±1.93)%,P<0.05],并下调TOPOⅠ和TOPOⅡmRNA及蛋白的表达。结论:ELE联合HTM可增强对Raji细胞增殖的抑制作用,下调TOPOⅠ、TOPOⅡ表达可能是其机制之一。

Objective：To observe the effects of elemene （ELE） combined with hyperthermia（HTM） on the proliferation and expression of topoisomerase Ⅰ and Ⅱ （ TOPOⅠ and Ⅱ ） in human Burkitt＇s lymphoma Raji cells. Methods：After treatment with ELE combined with HTM, cell proliferation was assessed by MTT, cell cycles were detected by FCM, TOPOI and TOPO Ⅱ mRNA were analyzed by RT- PCR,and TOPOI and TOPO Ⅱ protein expression were analyzed by Western Blot. Results： Compared to ELE alone and HTM alone, ELE combined with HTM showed a stronger inhibition to Raji cells proliferation in the dosage - and time -dependent ways （P 〈 0.05）, a lower IC50 （116 μg/ ml,84μg/ml,60μg/ml VS 93μml,68μml,41μg/ml） when compared to ELE alone at 24h,48h and 72h, a more marked tumor cells arrest at the S-phase [ （35.40±1.27）% ,（42.38±2.50） % VS （48.88 ±1.93）%, P 〈 0. 05 ], and a lower expressions of mRNA and protein for TOPOI and TOPO II were observed. Conclusion： ELE combined with HTM enhances the inhibition to the proliferation of Raji cells, which underlying mechanism might be related to down - regulated expression of topoisomerase Ⅰ and Ⅱ.